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Analysis of the puc Operon Promoter from Rhodobacter capsulatus

机译:荚膜红杆菌puc操纵子启动子的分析

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摘要

Expression of the Rhodobacter capsulatus puc operon, which codes for structural polypeptides of the light-harvesting-II peripheral antenna complex, is highly regulated in response to alterations in oxygen tension and light intensity. To obtain an understanding of the puc promoter region we report the high-resolution 5′ mapping of the puc mRNA transcriptional start site and DNA sequence analysis of the puc upstream regulatory sequence (pucURS). A ς70-type promoter sequence was identified (pucP1) which has a high degree of sequence similarity with carotenoid and bacteriochlorophyll biosynthesis promoters. Inspection of the DNA sequence also indicated the presence of two CrtJ and four integration host factor (IHF) binding sites. Transcriptional fusions of the pucURS fused to lacZ also confirmed that puc promoter activity is regulated by the transcriptional regulators IHF, CrtJ, and RegA. Gel retardation analysis using cell extracts indicates that mutations in IHF and RegA disrupt protein binding to DNA fragments containing the pucURS.
机译:响应于氧气张力和光强度的变化,高度编码调节荚膜红球菌操纵子的表达,该表达编码光捕获II外围天线复合体的结构多肽。为了了解puc启动子区域,我们报道了puc mRNA转录起始位点的高分辨率5'定位以及puc上游调控序列(pucURS)的DNA序列分析。鉴定出一个 70 型启动子序列(pucP1),该序列与类胡萝卜素和细菌叶绿素生物合成启动子具有高度的序列相似性。对DNA序列的检查还表明存在两个CrtJ和四个整合宿主因子(IHF)结合位点。 pucURS与lacZ融合的转录融合体也证实puc启动子活性受转录调节剂IHF,CrtJ和RegA调节。使用细胞提取物的凝胶阻滞分析表明,IHF和RegA中的突变破坏了蛋白质与包含pucURS的DNA片段的结合。

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